Mouse Tessp1 has been shown to be a testis-specific gene that may contribute to spermatogenesis. In this report, we raised a specific antibody against TESSP1 to assess its biological role. Western blotting detected testicular TESSP1 in all postnatal developmental stages of the mouse. Experiments using the testes of W/W^v mice, which lack germ cells, indicated TESSP1 expression in Sertoli cells and Leydig cells. In immunofluorescence staining of the wild-type mouse testis, dot-like signals for TESSP1 were observed in the adluminal compartment of the seminiferous tubules, while diffused signals were found in the basal compartment. Generally, the dot-like and diffused signals overlapped with the trans-Golgi network marker RAB6 and the transmembrane protein CADHERIN 2, respectively. Some TESSP1 staining was also observed in association with interstitial Leydig cells of the testis. The results of this study suggest that TESSP1 is predominantly localized in the plasma membrane of spermatogonia and Sertoli cells in the basal compartment, but exhibits an intracellular localization, presumably in the Golgi apparatus, of spermatocytes and spermatids in the adluminal compartment of the seminiferous tubules. The expression of TESSP1 in both germ cells and somatic cells and alteration in its cellular localization in the germ cells during spermatogenesis indicate that it may have a unique role in the testis.